Expression and functional verification of autophagy genes in <i>Chlamydomonas</i> and <i>Chlorella</i>.

<p>A: RT-PCR analysis of selected proteins in <i>Chlamydomonas</i> and <i>Chlorella</i> to confirm the sequences obtained from in silico research. Total RNA samples were collected during autophagy as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041826#s4" target="_blank">material and methods</a> section. Data are representative of three independent experiments. B: 3-MA (PI3K pathway inhibitor) blocks the formation of autophagic vacuole in <i>Chlorella</i>. Left: Autotrophically grown <i>Chlorella</i> cell resuspended in the heterotrophic medium and sampled after 6 h. Autophagic vacuoles disperse throughout the cytoplasm. Right: Autotrophically grown <i>Chlorella</i> cell resuspended in the heterotrophic medium for 6 h with 3-MA. There are no obvious autophagic vacuoles. The bar represents 500 nm. White arrowhead: lipid body; black arrowhead: autophagic vacuole. Data represent three independent experiments (A total of 208 control cells/201 test cells were analyzed).</p

Distribution of ATG proteins and domains in microalgae and <i>M. brevicollis</i>: ATG proteins exclusively involved in pexophagy.

<p>Na, not present or not identifiable; numbers indicate GenBank DNA accession numbers.</p

Distribution of ATG proteins and domains in microalgae and <i>M. brevicollis</i>: ATG proteins involved in PI3K complex.

<p>Na, not present or not identifiable; numbers indicate GenBank DNA accession numbers.</p

Domain organization of putative ATG17 proteins.

<p>Species and sequences are as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041826#pone-0041826-t001" target="_blank">Table 1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041826#pone-0041826-t007" target="_blank">Table 7</a>. Fused domains that form a single polypeptide chain are connected by a horizontal line. Figures are not drawn to scale.</p

Distribution of ATG proteins and domains in microalgae and <i>M. brevicollis</i>: Other ATG proteins pexophagy.

<p>Na, not present or not identifiable; numbers indicate GenBank DNA accession numbers.</p

Distribution of ATG proteins and domains in microalgae and <i>M. brevicollis</i>: ATG proteins involved in ATG9-Cycling system.

<p>Na, not present or not identifiable; numbers indicate GenBank DNA accession numbers.</p

Distribution of ATG proteins and domains in microalgae and <i>M. brevicollis</i>: Non-yeast ATG proteins.

<p>Distribution of ATG proteins and domains in microalgae and <i>M. brevicollis</i>: Non-yeast ATG proteins.</p

Distribution of ATG proteins and domains in microalgae and <i>M. brevicollis</i>: ATG proteins involved in Ubiquitin-Like Protein Conjugation Systems.

<p>Na, not present or not identifiable; numbers indicate GenBank DNA accession numbers.</p

Distribution of autophagy components in microalgae and choanoflagellate.

<p>Blank box: no homologues or putative orthologs could be found in any of the seven microalgae genomes; green box: predicted orthologs were detected in each of the genomes studied; blue box: putative orthologs were detected in some algal genomes; yellow box: homologues could only be found in <i>Monosiga brevicollis</i>. A: Distribution of putative ATG proteins in “core autophagic machinery”. B: The existence of homologues of non-yeast ATG proteins. C: The existence of the orthologs of additional pathway-specific requirement.</p

Species analyzed in this study.

<p>Na, not present.</p